6RG0

Structure of pdxj

6RG0 の概要

• エントリーDOI: 10.2210/pdb6rg0/pdb
• 分子名称: Pyridoxine 5'-phosphate synthase (1 entity in total)
• 機能のキーワード: serine phosphatase serb, serine aminotransferase serc, histidinol phosphatase hisb, histidinol phosphatase aminotransferase hisc, transferase
• 由来する生物種: Escherichia coli K12
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 116906.72

構造登録者

Rohweder, B.,Rajendran, C.,Sterner, R. (登録日: 2019-04-16, 公開日: 2020-05-13, 最終更新日: 2024-01-24)

主引用文献

Rohweder, B.,Lehmann, G.,Eichner, N.,Polen, T.,Rajendran, C.,Ruperti, F.,Linde, M.,Treiber, T.,Jung, O.,Dettmer, K.,Meister, G.,Bott, M.,Gronwald, W.,Sterner, R.
Library Selection with a Randomized Repertoire of ( beta alpha )8-Barrel Enzymes Results in Unexpected Induction of Gene Expression.
Biochemistry, 58:4207-4217, 2019

PubMed Abstract: The potential of the frequently encountered (βα)-barrel fold to acquire new functions was tested by an approach combining random mutagenesis and selection . For this purpose, the genes encoding 52 different phosphate-binding (βα)-barrel proteins were subjected to error-prone PCR and cloned into an expression plasmid. The resulting mixed repertoire was used to transform different auxotrophic strains, each lacking an enzyme with a phosphate-containing substrate. After plating of the different transformants on minimal medium, growth was observed only for two strains, lacking either the gene for the serine phosphatase SerB or the phosphoserine aminotransferase SerC. The same mutants of the genes E (encoding a putative -acetylmannosamine-6-phosphate 2-epimerase) and J (encoding the pyridoxine 5'-phosphate synthase) were responsible for rescuing both ΔB and ΔC. Unexpectedly, the complementing NanE and PdxJ variants did not catalyze the SerB or SerC reactions . Instead, RT-qPCR, RNAseq, and transcriptome analysis showed that they rescue the deletions by enlisting the help of endogenous enzymes HisB and HisC through exclusive up-regulation of histidine operon transcription. While the promiscuous SerB activity of HisB is well-established, our data indicate that HisC is promiscuous for the SerC reaction, as well. The successful rescue of ΔB and ΔC through point mutations and recruitment of additional amino acids in NanE and PdxJ provides another example for the adaptability of the (βα)-barrel fold.

PubMed: 31557000
DOI: 10.1021/acs.biochem.9b00579

実験手法

X-RAY DIFFRACTION (3.074 Å)